Heinzer Stefan, Krucker Thomas, Stampanoni Marco, Abela Rafael, Meyer Eric P, Schuler Alexandra, Schneider Philipp, Müller Ralph
Institute for Biomedical Engineering, University and ETH Zürich, Zürich, Switzerland.
Neuroimage. 2006 Aug 15;32(2):626-36. doi: 10.1016/j.neuroimage.2006.03.043. Epub 2006 May 11.
There is a wide range of diseases and normal physiological processes that are associated with alterations of the vascular system in organs. Ex vivo imaging of large vascular networks became feasible with recent developments in microcomputed tomography (microCT). Current methods permit to visualize only limited numbers of physically excised regions of interests (ROIs) from larger samples. We developed a method based on modified vascular corrosion casting (VCC), scanning electron microscopy (SEM), and desktop and synchrotron radiation microCT (SRmicroCT) technologies to image vasculature at increasing levels of resolution, also referred to as hierarchical imaging. This novel approach allows nondestructive 3D visualization and quantification of large microvascular networks, while retaining a precise anatomical context for ROIs scanned at very high resolution. Scans of entire mouse brain VCCs were performed at 16-microm resolution with a desktop microCT system. Custom-made navigation software with a ROI selection tool enabled the identification of anatomical brain structures and precise placement of multiple ROIs. These were then scanned at 1.4-microm voxel size using SRmicroCT and a local tomography setup. A framework was developed for fast sample positioning, precise selection of ROIs, and sequential high-throughput scanning of a large numbers of brain VCCs. Despite the use of local tomography, exceptional image quality was achieved with SRmicroCT. This method enables qualitative and quantitative assessment of vasculature at unprecedented resolution and volume with relatively high throughput, opening new possibilities to study vessel architecture and vascular alterations in models of disease.
有多种疾病和正常生理过程与器官血管系统的改变有关。随着微计算机断层扫描（microCT）技术的最新发展，大型血管网络的离体成像变得可行。目前的方法只能可视化从较大样本中物理切除的有限数量的感兴趣区域（ROI）。我们开发了一种基于改良血管铸型（VCC）、扫描电子显微镜（SEM）以及台式和同步辐射微CT（SRmicroCT）技术的方法，以在不断提高的分辨率水平下对血管系统进行成像，也称为分层成像。这种新方法允许对大型微血管网络进行无损三维可视化和定量分析，同时为以非常高分辨率扫描的ROI保留精确的解剖背景。使用台式微CT系统以16微米的分辨率对整个小鼠脑VCC进行扫描。带有ROI选择工具的定制导航软件能够识别脑解剖结构并精确放置多个ROI。然后使用SRmicroCT和局部断层扫描装置以1.4微米的体素大小对这些ROI进行扫描。开发了一个框架，用于快速样本定位、精确选择ROI以及对大量脑VCC进行顺序高通量扫描。尽管使用了局部断层扫描，但SRmicroCT仍获得了出色的图像质量。该方法能够以前所未有的分辨率和体积对血管系统进行定性和定量评估，且通量相对较高，为研究疾病模型中的血管结构和血管改变开辟了新的可能性。
